The Cloning and Expression of the Recombinant Transcriptional Activator AFT1 From Saccharomyces cerevisiae
School Name
South Carolina Governor's School for Science & Mathematics
Grade Level
12th Grade
Presentation Topic
Biochemistry
Presentation Type
Mentored
Abstract
To better understand Aft1's (Activator of Ferrous Transport 1) molecular mechanism of iron regulation in baker's yeast (Saccharomyces cerevisiae), our lab has attempted to express and purify the full length Aft1 protein in E. coli for biochemical analysis. However, these previous attempts have failed. During my six weeks of research, we have successfully constructed an expression vector containing the truncated version of the Aft1 gene using the Gibson assembly protocol. This new plasmid will allow for the expression and purification of Aft1. With large amounts of the purified protein, our lab will be able to explore the DNA and iron-binding properties of Aft1 at the molecular level, allowing for a better understanding of how the protein regulates iron.
Recommended Citation
Benson, Abigail, "The Cloning and Expression of the Recombinant Transcriptional Activator AFT1 From Saccharomyces cerevisiae" (2020). South Carolina Junior Academy of Science. 278.
https://scholarexchange.furman.edu/scjas/2020/all/278
Location
Furman Hall 118
Start Date
3-28-2020 10:45 AM
Presentation Format
Oral Only
Group Project
No
The Cloning and Expression of the Recombinant Transcriptional Activator AFT1 From Saccharomyces cerevisiae
Furman Hall 118
To better understand Aft1's (Activator of Ferrous Transport 1) molecular mechanism of iron regulation in baker's yeast (Saccharomyces cerevisiae), our lab has attempted to express and purify the full length Aft1 protein in E. coli for biochemical analysis. However, these previous attempts have failed. During my six weeks of research, we have successfully constructed an expression vector containing the truncated version of the Aft1 gene using the Gibson assembly protocol. This new plasmid will allow for the expression and purification of Aft1. With large amounts of the purified protein, our lab will be able to explore the DNA and iron-binding properties of Aft1 at the molecular level, allowing for a better understanding of how the protein regulates iron.
